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ISSN: 1579-4377

LAWSONIA INERMIS IN THE CONTROL OF MELOIDOGYNE SPP. ON


CORCHORUS OLITORUS
1
Fabiyi Oluwatoyin Adenike and 2Atolani Olubunmi*.
1
Department of Crop Protection, University of Ilorin, Ilorin, Nigeria
2
*Department of Chemical Sciences, Redeemer’s University,
P.M.B.3005, Redemption Camp. Mowe, Ogun State, Nigeria.
tolanvent@yahoo.com

ABSTRACT

The efficacy of cypress shrub, Lawsonia inermis L. in the control of root – knot nematodes,
Meloidogyne spp was investigated in the laboratory and the Screenhouse. In the laboratory, only the
aqueous extract of the test plant (Lawsonia inermis) was assessed for its effects on nematode egg
hatch and juvenile mortality. For the Screenhouse, the experimental design was a 2x2x3 factorial
experiment, fitted into a randomized complete block design, involving two levels of nematode
population (0 and 300) and two forms of test plant application (aqueous extract and powdered form).
In the laboratory, the aqueous extracts of Lawsonia inermis caused a significant reduction (P< 0.05)
in nematode egg hatched. Egg hatch was 92.0% in the untreated control (0% extract) as compared to
11.7% in 25% crude extract of Lawsonia inermis (L) extract. Within two days of the experiment
98.4% mortality was observed in 100% crude concentration of Lawsonia inermis (L), as compared
with 0% mortality in the control. All concentrations of the extract caused significantly higher
mortality than the control. In the screenhouse, 15% aqueous crude concentration, significantly
reduced nematode population in the root and in the soil. All treated plants were also less galled
compared to the untreated control.

KEYWORDS

Lawsonia inermis, Meloidogyne spp, nematodes, screenhouse.


Olubunmi, A.  et al. EJEAFChe, 10 (3), 2011. [2000-2006]

INTRODUCTION

Jew’s mallow, Corchorus olitorius (L) belongs to the family Tiliaceae, it is of utmost importance in
the south western part of Nigeria where it is widely consumed, very popular and highly appreciated.
Corchorus olitorius has high moisture content in its fresh form with considerable quantities of
vitamins which help protect the body against diseases and contribute in no small measure to good
health (Agusiobo, 1984).
Vegetables compensate for the scarce animal protein sources in rural communities in Nigeria
(Oyenuga and Fetuga, 1975). The Asian vegetable Research and Development Center in its strategic
plan1991 reported that vegetable production offers great income to small scale farmers and low
income earners. It provides employment opportunities for women, hence it is an important revenue
source in Nigeria (AVRDC, 1991).
However, plant parasitic nematodes are recognized as one of the major pests of C. olitorius in
the tropics. The most destructive nematodes responsible for enormous yield loss in C. olitorius are
the root – knot nematodes, Meloidgyne spp. The disease inflicted by this group of nematodes is
characterized by numerous pronounced swellings or galls on the root of susceptible host plant with
consequent reduction in growth and yield of infected plants.
Synthetic chemicals constituted the world most effective nematode control weapon put in
place by man, because of its quick efficacy. This strategy ruled the world of nematode control for
more than ten decades (Johnson, 1987). However nematicides are expensive and hazardous because
of the residual effect on the crop, soil and non target organisms.
As a result of the side effects and high costs associated with the use of synthetic nematicides,
efforts have been made to study the nematicidal potentials of some plants on plant parasitic nematode
Khan and Saxena (1997) in their experiment on the integrated management of root knot nematode, M.
javanica infecting tomato using amendments of oil cakes, bone and horn meals, found that there was
increase in growth of tomato and reduction in nematode multiplication.Kimpinski et al (2000)
reported that Tagetes tennifolia, T. patula and T. erecta were found to significantly reduce the
population of Pratylenchus penetrans on potato, Pandey (2000) observed maximum reduction in
M.incognita populations at high doses of three Organic nematicides. Oyedunmade et al (2000)
reported that neem leaf extract had nematicidal potential, and caused an improvement in the growth
and yield of okro.
Oyedunmade (2004) also reported that African marigold, Tagetes erecta has nematicicidal
properties with positive effect on the vegetative growth of treated plant in the field. However,
there are still several natural materials whose potentials remain untapped. This study was therefore
conducted with the following objectives in mind.
(1) To determine the active chemical components present in Lawsonia inermis (2) To
compare nematicidal properties of the aqueous extract and powdered form of test plant. L. inermis on
the growth and yield of C. olitorius (3) To compare the effects of the different concentrations of the
aqueous extract of L. inermis (L) on the survival of root knot nematode juveniles and eggs in the
laboratory.

2001
Olubunmi, A.  et al. EJEAFChe, 10 (3), 2011. [2000-2006]

MATERIALS AND METHODS

Source and Preparation of Test plant extract:-


The test plant, Lawsonia inermis was collected from Ile apa village, close to the Faculty of
Agriculture, University of Ilorin. It was air-dried thoroughly until brittle. The leaves were removed
from the stem after drying and pound in mortar into powdered form.
One kilogram of powdered plant material was weighed and soaked in a liter of water for 24 hours,
thereafter filtered, and the filtrate concentrated to 100 ml by heating on the water bath to remove
excess water. The concentrated plant extract was taken as 100% concentration, from this, serial
dilutions were made to obtain 25, 50 and 75%, extracts. Ordinary water served as the 0% extract
(control).

Nematode egg hatch and juvenile mortality trials:-

Twenty (20) Petri dishes containing 8 ml of each concentration of plant extract and 5 Petri dishes
containing 8 ml of water were used for the juvenile mortality assessment and egg hatch ability.
For the Juvenile mortality experiment, 300 juveniles were introduced into each Petri dish.
Counting of nematodes was done under the stereo microscope at 2hours, 4hours and daily for seven
days. Those juveniles that did not respond to the touch of a needle were considered dead. For the egg
hatch test 300 eggs were introduced into each Petri dish. The Petri dishes were covered to prevent
evaporation while count of hatched eggs was done at 24 hours intervals for ten (10) consecutive days
under the stereo microscope.

Potted plant experiment:-

Soil Sterilization:- Sandy loam soil was collected behind the Faculty of Agriculture, University of
Ilorin. The soil was sieved to remove gravels, stone and plant debris with a 2mm mesh sieve. The
soil was put in a metal drum and then pasteurized for 4 hours at 60 oC (Gautam and Goswami, 2002).
After cooling, the soil was weighed into experimental pots at 10kg per pot and the pots were placed
on bricks to avoid microbial infestation in the course of the experiment.

Source and Extraction of Nematode Juveniles:

Galled roots of Corchorus olitorius was collected from Rice Research, Seed Multiplication and
Demonstration Farm, Sango Ilorin: The roots were taken to the laboratory, washed gently under
running tap to remove attached soil, small stones and other plant debris. The roots were later cut with
scissors into small pieces and macerated in a warring blender for 30 seconds only. The blended
mixture was then poured over a bank of sieves that had bean arranged in order of mesh size viz,
10mm, 100mm, and 200mm. The contents of the 200mm mesh were carefully collected and rinsed
into a beaker with little water. Whitehead and Hemming (1965) tray method of nematode extraction
was used.

Planting Operation: - The seeds were planted at a sowing depth of 0.3cm, in the little holes made in
the center of the sterilized soil in the experimental pots, and covered with soil. The resulting
seedlings were watered daily. Germination of seeds started on the fourth day, and the plants were
thinned down to a plant per pot by removing excess plants on the 10th day after germination.

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Olubunmi, A.  et al. EJEAFChe, 10 (3), 2011. [2000-2006]

Nematode inoculation and treatment application: After the 14th day of planting 300 juveniles of
Meloidogyne spp were inoculated close to the roots of Corchorus olitorius plant in each of thirty pots
while another set of thirty other pots were not inoculated. Seven days after inoculation, the
inoculated and uninoculated pots were treated with powdered form and aqueous extract of Lawsonia
inermis, at various rates, close to the root of the plants. The rates were 0, 15 and 30g/pot for the
powdered form of treatment, while for the aqueous form, the rates used were 0 , 7.5 and 15%.

Data collection and statistical analysis: - Data were collected weekly from the experimental plants
to record the following parameters: Plant height, number of leaves per plant and number of branches
per plant. Shoot weight, root weight, root gall index (Taylor & Sasser, 1978), nematode population
per 250g soil sample and nematode population in 5g root sample were assessed at plant harvest. All
data from the experiment were subjected to analysis of variance (ANOVA). Treatment means were
separated using the new Duncan’s Multiple Range Test at 5% level of probability.

RESULTS AND DISCUSSION

Phytochemical screening: Tannin, saponin, alkaloid, glucoside, anthraquinone and flavonoids were
found present in the test plant (L. inermis)
Laboratory tests: - The effect of the different concentration of aqueous extracts of Lawsonia
inermis on the egg hatch of Meloidogyne spp is shown in Table 1. The plant extracts were effective
in inhibiting nematode egg hatch, as all the concentrations of Lawsonia inermis used were
significantly more effective than the control in inhibiting egg hatch, although the lowest
concentration was not as effective as the other higher concentrations. The cumulative egg hatch in
the control was 92% at the end of the experiment while the highest percentage cumulative egg hatch
observed was 11.7% in 25% extract treatment. The higher concentrations of 50, 75, and 100%
aqueous extract of Lawsonia inermis did not record any egg hatch. The control experiment recorded
an egg hatch value of 14% and 90.5% at day 1 and day 9 respectively.

TABLE 1:- Effects of different concentrations of aqueous extract of Lawsonia inermis on percentage egg hatch of
meloidogyne spp.

Lawsonia inermis DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 DAY 8 DAY 9 DAY 10
0 Control 14.06 17.3c 37.3c 40.56 49.2c 62.8c 76.5c 80.0c 90.5c 92.0c
25% Crude Extract 0.6a 2.4b 2.8b 4.0b 4.9b 6.3b 9.3b 10.1b 11.1b 11.7b
50% Crude Extract 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a
75% Crude Extract 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a
100% Crude Extract 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a
Standard Error 0.179 0.190 0.536 0.553 1.460 0.298 0.190 0.378 0.104 0.162
Means in given column followed by the same letter are not significantly different at P< 0.05 using the new Duncan’s
Multiple Range Tests.

Between two and four hours of the exposure of juveniles to test plant extract, there was no
significant difference in the effect of all the levels of concentration of Lawsonia inermis on juvenile
mortality (Table 2). At six hours however, there were significant differences in juvenile mortality and
the 100% concentration of extract caused the highest mortality of 26.6%. Percentage mortality
ranged between 24.8 and 93% at day 1 of the experiment. The control experiment had the lowest
percentage mortality during the experimental period. The first record of juvenile mortality of 2.1%

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Olubunmi, A.  et al. EJEAFChe, 10 (3), 2011. [2000-2006]

was observed in the control treatment on the 4th day of the experiment. This juvenile mortality in the
control only increased to 6.0 on the 7th day of the experiment. The level of juvenile mortality
increased with increase in the exposure period. At the end of the experiment, an average of 100%
mortality was observed in all the concentrations of test plant extract as compared with the control
experiment which had 6.0%, mortality.

Table 2:- Effects of different concentrations of aqueous extract of lawsonia inermis on percentage junenile mortality of
meloidogyne spp. Exposure time of juveniles to extract

Lawsonia inermis 2 Hrs 4 Hrs 6 Hrs 8 Hrs DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 DAY
0 Control 0.0 0.0 0.0c 0.0c 0.0c 0.0c 0.0d 2.1c 3.3c 7
25% Crude extract 0.0 0.0 3.2d 3.2d 24.8d 40.8d 58.2c 73.66 89.8b 6.0b
50% Crude extract 0.0 0.0 6.7c 7.3c 48.8c 67.2c 86.2b 97.4a 100.0a 98.6a
75% Crude extract 0.0 0.0 15.2b 16.3b 78.4b 83.8b 97.6a 99.6a 100.0a 100.0a
100% Crude extract 0.0 0.0 26.6a 28.6a 93.0a 98.4a 100.0a 100.0a 100.0a 100.0a
Standard error 0.142 N.S 0.277 N.S 0.529 0.515 1.465 1.390 1.995 1.614 1.190 100.0a
Means in a given column followed by the same letter are not significantly different at P < 0.05 using the new Duncan’s Multiple Range
Test. N.S: Not Significant.

Table 3: Main effect of nematode inoculation, forms of application and level of application of lawsonia inermison, on
different growth parameters of corchorus olitorius

Number of branches
Plant height Number of leaves
13 13 7 10 13
Inoculation 7 WAP 10 WAP 7 WAP 10 WAP
WAP WAP WAP WAP WAP
Not
inoculate(0 39.7a 70.6a 110.9a 49.4a 88.3a 134.7a 9.1a 15.3a 23.3a
juvenile)
Inoculated
25.9b 37.9b 53.4b 226.8b 46.0b 66.0b 4.8b 8.0b 11.7b
(300 juvenile)
S.E 0.516 0.493 0.921 0.651 1.056 0.870 0.237 0.316 0.350
Form
Aqueous 32.7 54.6 92.2a 40.4a 75.6a 117.1a 7.8a 13.8a 19.8a
Powder 32.9 53.8 82.1 35.8b 58.8b 83.6b 6.1b 9.5b 15.2b
S.E 0.516N.S 0.493N.S 0.921 0.651 11.05b 0.870 0.237 0.316 0.350
Level
0 Control 28.3c 47.3b 70.4c 31.0b 51.3b 69.6c 5.1c 8.8c 14.5c
7.5 Conc 15g
34.1b 57.4a 86.2b 41.5a 73.6a 113.2b 6.7b 10.5b 16.4a
/10kg soil
15% Conc
30g / 10Kg 36.1a 50.2b 89.8a 41.9a 76.6a 118.3a 9.2a 15.7a 21.7a
soil
S.E 0.632 0.499 128 00.797 11.294 1.066 0.290 0.428
Means in a given column followed by the same letter are not significantly different at
P < 0.05 using the new Duncan’s Multiple Range Test. N.S: Not Significant.

Pot Experiment:
Nematode inoculation resulted in a significant reduction in the vegetative growth of Corchorus
olitorius, as plant height, number of leaves, and number of branches per plant were significantly
higher in uninoculated plants (Table 3). The test plant, Lawsonia inermis significantly reduced the
effect of the nematodes on Corchorus olitorius, as treated plants had better vegetative growth than
the untreated ones. The higher doses of treatment seemed to be more effective than the lower doses.

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Olubunmi, A.  et al. EJEAFChe, 10 (3), 2011. [2000-2006]

While the aqueous form of treatment was significantly better than the powdered form in
increasing the plant height, number of leaves, and number of branches per plant.

Table 4:- The main effect of nematodes inoculation forms of application and level of application of Lawsonia inermis on
root gall rating and nematode populations in soil sample or Corhorus olitorius roots.

Nematode population in 250g Nematode population in 5g


Inoculation Root Gall Rating
soil sample root sample
Not Inoculation
0.0b 0.0b 0.0b
(0 Juvenile)
Inoculation
241.2a 128.4a 1.2a
(300 Juveniles)
S.E 0.651 3.167 0.00
Form
Aqueous 118.7b 61.1 0.7b
Powder 122.4a 67.3 1.0a
S.E 0.651 3.167 NS 0.00
Level
0 (Control) 355.1a 179.5a 2.0a
7.5 CONC 15g /10kg soil 6.7b 13.1b 0.5b
15% CONC 30g / 10Kg soil 0.0c 0.0c 0.0c
S.E 0.798 3.878 0.00
Means in the same column followed by the same letter are not significantly different at P> 0.05 using the New Duncan’s Multiple
Range Test. N.S – Not Significant

Furthermore, nematode inoculation resulted in a significant increase in nematode population


in 250g soil sample, nematode population in root sample, and root gall rating (Table 4). The aqueous
form of Lawsonia inermis, however significantly reduced nematode population in 5g root sample,
250g soil sample, and root gall rating. The higher doses of treatment were very effective as there was
no nematode in 250g soil sample and 5g root sample, and consequently no root galling (Table 4).

DISCUSSION

The bio-active compounds (tannin, saponin, flavonoids, glucosides, alkaloids and anthraquinone
present in the test plant could have been the reason for the efficacy of Lawsonia inermis in
controlling root-knot nematodes in Corchorus olitorius. The observed nematotoxic effects Lawsonia
inermis may be attributed to the presence, in the plant, of nematicide chemical compound that are
injurious to the eggs and infective second stage juveniles of Meloidogyne spp. Deverall (1972)
reported the pesticidal astringency and phyto-repellent nature of flavonoids and other bioactive
compounds present in plants. Paulo (2001) reported the efficacy of the aqueous extracts of the roots
and shoots of Artemisia annua, which at concentration of 100% resulted in 100% Meloidogyne
incognita juvenile mortality within 24 hours of exposure in vitro.
Apart from the nematicidal properties of the plant material, it is possible for the plant powder
and extract to increase the nutrient status of the soil and encourage better crop growth and yield.
Gautam and Goswami (2002) reported improved yield in a specie of cowpea when neem cake was
used in the treatment of Meloidogyne incognita. The ready availability of Lawsonia inermis and its
effect on nematode population and plant growth suggest interests in its potentials as a biopesticidal
product for the control of plant – parasitic nematode.

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